Consequences of zygote injection and germline transfer of mutant human mitochondrial DNA in mice

Hong Yu, Rajeshwari D. Koilkonda, Tsung Han Chou, Vittorio Porciatti, Arpit Mehta, Ian D. Hentall, Vince A. Chiodo, Sanford L. Boye, William W. Hauswirth, Alfred S. Lewin, John Guy

Research output: Contribution to journalArticlepeer-review

21 Scopus citations


Considerable evidence supports mutations in mitochondrial genes as the cause of maternally inherited diseases affecting tissues that rely primarily on oxidative energymetabolism, usually the nervous system, the heart, and skeletal muscles. Mitochondrial diseases are diverse, and animal models currently are limited. Here we introduced a mutant human mitochondrial gene responsible for Leber hereditary optic neuropathy (LHON) into the mouse germ line using fluorescence imaging for tissue-specific enrichment in the target retinal ganglion cells. A mitochondria-targeted adeno-associated virus (MTS-AAV). containing the mutant human NADH ubiquinone oxidoreductase subunit 4 (ND4) gene followed by mitochondrial-encoded mCherry was microinjected into zygotes. Female founders with mCherry fluorescence on ophthalmoscopywere backcrossed with normal males for eight generations. Mutant human ND4 DNA was 20% of mouse ND4 and did not integrate into the host genome. Translated human ND4 protein assembled into host respiratory complexes, decreasing respiratory chain function and increasing oxidative stress. Swelling of the optic nerve head was followed by progressive demise of ganglion cells and their axons, the hallmarks of human LHON. Early visual loss that began at 3 mo and progressed to blindness 8 mo after birth was reversed by intraocular injection of MTS-AAV expressing wild-type human ND4. The technology of introducing human mitochondrial genes into themouse germ line has never been described, to our knowledge, and has implications not only for creating animal models recapitulating the counterpart human disorder but more importantly for reversing the adverse effects of the mutant gene using gene therapy to deliver the wild-type allele.

Original languageEnglish (US)
Pages (from-to)E5689-E5698
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number42
StatePublished - Oct 20 2015


  • Adeno-associated virus
  • Blindness
  • Gene therapy
  • Leber hereditary optic neuropathy
  • Mitochondria

ASJC Scopus subject areas

  • General


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